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杜氏利什曼原虫核糖体 P1 基因作为 DNA 疫苗在实验性内脏利什曼病中的疗效。

Efficacy of Leishmania donovani ribosomal P1 gene as DNA vaccine in experimental visceral leishmaniasis.

机构信息

Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India.

出版信息

Exp Parasitol. 2011 Sep;129(1):55-64. doi: 10.1016/j.exppara.2011.05.014. Epub 2011 May 27.

Abstract

The acidic ribosomal proteins of the protozoan parasites have been described as prominent antigens during human disease. We present here data showing the molecular cloning and protective efficacy of P1 gene of Leishmania donovani as DNA vaccine. The PCR amplified complete ORF cloned in either pQE or pVAX vector was used either as peptide or DNA vaccine against experimentally induced visceral leishmaniasis in hamsters. The recombinant protein rLdP1 was given along with Freund's adjuvant and the plasmid DNA vaccine, pVAX-P1 was used alone either as single dose or double dose (prime and boost) in different groups of hamsters which were subsequently challenged with a virulent dose of 1×10(7) L. donovani (MHOM/IN/DD8/1968 strain) promastigotes by intra-cardiac route. While the recombinant protein rLdP1 or DNA vaccine pVAX-P1 in single dose format were not found to be protective, DNA vaccine in a prime-boost mode was able to induce protection with reduced mortality, a significant (75.68%) decrease in splenic parasite burden and increased expression of Th1 type cytokines in immunized hamsters. Histopathology of livers and spleens from these animals showed formation of mature granulomas with compact arrangement of lymphocytes and histiocytes, indicating its protective potential as vaccine candidate.

摘要

原生动物寄生虫的酸性核糖体蛋白在人类疾病中被描述为主要抗原。我们在此介绍了利什曼原虫 DNA 疫苗的 P1 基因的分子克隆和保护效力的数据。PCR 扩增的完整 ORF 克隆在 pQE 或 pVAX 载体中,无论是作为肽还是 DNA 疫苗,都可用于对抗仓鼠实验性内脏利什曼病。重组蛋白 rLdP1 与福氏佐剂一起给药,而质粒 DNA 疫苗 pVAX-P1 单独使用,无论是单次剂量还是双剂量(一次接种和加强接种),都用于不同组的仓鼠中,随后通过心内途径用 1×10(7)L. donovani (MHOM/IN/DD8/1968 株) 前鞭毛体的强毒剂量进行攻击。虽然重组蛋白 rLdP1 或 DNA 疫苗 pVAX-P1 的单次剂量形式不能提供保护,但 DNA 疫苗的一次接种和加强接种模式能够诱导保护,降低死亡率,脾脏寄生虫负荷显著(75.68%)降低,免疫仓鼠中 Th1 型细胞因子的表达增加。这些动物的肝脏和脾脏组织病理学显示成熟肉芽肿的形成,淋巴细胞和组织细胞排列紧凑,表明其作为候选疫苗具有保护潜力。

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